HIF-1α基因表达沉默对肝癌细胞的放射增敏作用  

Effect of Silencing HIF-1α on Radiosensitization in Human Hepatocellular Carcinoma Cell Line

作　　者：吴德华[1] 朱晓霞[1] 张洪波[1] 陈龙华[1] 

机构地区：[1]南方医科大学南方医院放疗科

出　　处：《中国肿瘤临床》2009年第2期110-113,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30500242);广东省自然科学基金资助(编号:05004743)

摘　　要：目的：探讨在乏氧条件下，HIF—1α在肝癌细胞中的表达，及HIF—1α表达沉默后对肝癌细胞的放射增敏作用。方法：利用Western bolt方法检测不同乏氧时相HIF—1α在肝癌细胞Hep3b中的表达。构建特异性的HIF—1α慢病毒干扰载体，利用慢病毒感染肝癌细胞Hep3b，杀稻瘟菌素进行药物筛选，利用RT—PCR和Western blot检测干扰效果。利用克隆形成实验观察HIF—1α表达沉默后对肝癌细胞反射敏感性的影响。结果：在乏氧条件下，肝癌Hep3b细胞HIF-1α蛋白表达增强。利用特异性的HIF-1α慢病毒沉默肝癌细胞HIF-1α的表达，Hep3b细胞中HIF—1α表达降低了90％，说明HIF—1α载体构建成功。HIF—1α表达沉默后，能明显降低放射后Hep3b细胞的克隆形成率，其放射增敏比为2．18。结论：乏氧可以诱导肝癌细胞HIF-1α的表达，HIF-1α表达沉默对肝癌细胞具有放射增敏作用。Objective： To study the expression of hypoxia inducible factor-1α （HIF-1α） in human hepatocellular carcinoma （HCC） cells under hypoxia and to explore the effect of silencing HIF-1α on radiosensitization in HCC. Methods： The expression of HIF-1α at different hypoxic culture phases was detected by Western blot. HIF1 a specific RNAi lentiviral vector was constructed. HIF-1α silencing cells were screened by blasticidin. RT-PCR and Western blot analysis were used to detect the expressions of HIF-1α mRNA and protein, respectively. Clonogenic assay was employed to observe its effects on the radiosensitivity after HIF-1α silencing. Resuits： Under hypoxia, the HIF-1α protein expression was increased. Transduction of HCC cells with HIF-1 a RNAi vector resulted in sequence specific silencing with decrease of HIF-1α expression by 90%. HIF-1α silencing significantly reduced the clonogenic activity and enhanced the radiosensitivity of Hep3b cells with a SER of 2.18. Conclusion： Hypoxia can induce HIF-1α expression. HIF-1α silencing can enhance the radiosensitivity of HCC.

关 键 词：HIF-1Α RNA干扰 放射增敏 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]